Process and apparatus for separation by liquid chromatography

Abstract

A separation process by liquid chromatography, which comprises sending a mixture sample in a large amount together with a liquid mobile phase to at least one column, into which a packing having a large particle size has been charged, to preliminarily fractionate the components of the mixture, taking out fractions containing objective components from the eluate from the first column, and recycling said fractions within a loop provided with at least one second column, into which a packing having a large particle size has been charged, to separate precisely the objective components. According to this process, it is possible to separate economically objective components from the sample loaded in a large amount.

What is claimed is: 
     
       1. A large-scale separating apparatus for separating an objective component from a large amount of a mixture sample by liquid chromatography, which has a preliminary fractionation line comprising: at least one first column, in which a first packing having a large particle size has been charged; an eluent tank provided on the upstream side of the first column; a sample injecting valve provided in a passage between the first column and the eluent bath; a sample bath communicated with the sample injecting valve; a first fraction collector provided through a passage on the downstream side of the first column; a first detector provided in a passage between the first column and the first fraction collector; a liquid transfer device provided in a liquid passage; and a pre-cut valve provided in a passage between the first column and the first fraction collector for taking out main fractions containing the objective component, and a recycling line comprising: at least one second column charged with a second packing having a large particle size, which column is to be communicated through a passage with the pre-cut valve in the aforementioned preliminary fractionation line when the main fractions are taken out; a second fraction collector provided through a passage on the downstream side of the second column; a second detector provided in a passage between the second column and the second fraction collector; a recycling valve for turning eluate from the second detector to the inlet of the second column upon recycling of the eluate and causing the objective component to flow into the second fraction collector after completion of separation; and a liquid transfer device provided in a passage between the recycling valve and the inlet of the second column; and wherein the sample injecting valve, the pre-cut valve and the recycling valve are changed over by respective controlling signals, and the operation states of the respective liquid transfer devices in the preliminary fractionation line and the recycling line are controlled according to respective controlling signals, which apparatus includes first and second fractionation valves which are respectively provided on the upstream sides of the first fraction collector and the second fraction collector and are changed over according to the respective signals for taking out samples, an operation controlling means from which controlling signals are sent out at predetermined times, a peak judging means for judging the peaks in the eluate by the detection signals respectively sent from the first detector and the second detector, and a controlling means for taking out the component which sends out taking out signals according to said judgments. 
     
     
       2. A process for separation by the apparatus as defined in claim 1 which comprises: loading the predetermined times and predetermined standards into the operation controlling means and the peak judging means, respectively in initial state of the apparatus;   operating the sample injecting valve, the pre-cut valve, the recycling valve and the liquid transfer devices in the preliminary fractionation line and the recycling line by respective controlling signals from the operation controlling means; and   judging the detecting signals of the peaks of eluates from the first detector and/or the second detector at the peak judging means based on the predetermined standards whereby, if the peak is not the one to be separated, the fraction corresponding to the peak is caused to flow to a drain without being taken out and, if the peak is the one to be separated, the fraction corresponding to the peak is taken out into a tank of the fraction collector.